lunes, 3 de noviembre de 2014

Development of an efficient entire capsid-coding region amplificati... - PubMed - NCBI

Development of an efficient entire capsid-coding region amplificati... - PubMed - NCBI

 2014 Oct 22. pii: JCM.02384-14. [Epub ahead of print]

Development of an efficient entire capsid-coding region amplification method for direct detection of poliovirus from stool extracts.


Laboratory diagnosis has played a critical role in the Global Polio Eradication Initiative (GPEI) since 1988 by isolating and identifying poliovirus (PV) from stool specimens by using cell culture, as a highly sensitive system to detect PV. In the present study, we aimed to develop a molecular method to detect PV directly from stool extracts with a high efficiency comparable to that of cell culture. We developed a method to efficiently amplify the entire capsid-coding region of human enteroviruses (EV) including PV. cDNAs of the entire capsid-coding region (3.9 kb) were obtained from as few as 50 copies of PV genomes. PV was detected from the cDNAs by an improved PV-specific real-time RT-PCR system and nucleotide sequence analysis of the VP1-coding region. For assay validation, we analyzed 84 stool extracts that were positive for PV in cell culture and detected PV genome from 100% of the extracts (84/84 samples) by this method in combination with a PV-specific extraction method. PV could be detected from 2/4 samples of stool extracts that were negative for PV in cell culture. In PV-positive samples, EV species C viruses were also detected with a high frequency (27%, 23/86 samples). This method would be useful for direct detection of PV from the stool extracts without using cell culture.
Copyright © 2014, American Society for Microbiology. All Rights Reserved.

[PubMed - as supplied by publisher]

No hay comentarios:

Publicar un comentario